#METABOLOMICS WORKBENCH imssr_20250602_212528 DATATRACK_ID:5983 STUDY_ID:ST003965 ANALYSIS_ID:AN006521 PROJECT_ID:PR002483 VERSION 1 CREATED_ON June 12, 2025, 10:27 am #PROJECT PR:PROJECT_TITLE Metabolomics of CAR-T cells and their products in leukemia patients PR:PROJECT_TYPE Untargeted metabolomics PR:PROJECT_SUMMARY Although most patients with relapsed/refractory B-cell acute lymphoblastic PR:PROJECT_SUMMARY leukemia (B-ALL) receiving CD19-targeted chimeric antigen receptor (CAR) T cell PR:PROJECT_SUMMARY therapy achieve remission, loss of CAR T cell functionality and subsequent PR:PROJECT_SUMMARY relapse remains an unmet therapeutic need. We applied an integrative approach to PR:PROJECT_SUMMARY study the immunometabolism of pre- and post-infusion CD19-CAR T cells of PR:PROJECT_SUMMARY patients with relapsed/refractory B-ALL. Pre-infusion CAR T cells of long-term PR:PROJECT_SUMMARY responders (LTR) had increased oxidative phosphorylation, fatty acid oxidation, PR:PROJECT_SUMMARY and pentose phosphate pathway activities, higher mitochondrial mass, tighter PR:PROJECT_SUMMARY cristae, and lower mTOR expression compared to products of short-term PR:PROJECT_SUMMARY responders. Post-infusion CAR T cells in bone marrow (BM) of LTR had high PR:PROJECT_SUMMARY immunometabolic plasticity and mTOR-pS6 expression supported by the BM PR:PROJECT_SUMMARY microenvironment. Transient inhibition of mTOR during manufacture induced PR:PROJECT_SUMMARY metabolic reprogramming and enhanced anti-tumor activity of CAR T cells. Our PR:PROJECT_SUMMARY findings provide insight into immunometabolic determinants of long-term response PR:PROJECT_SUMMARY and suggest a therapeutic strategy to improve long-term remission. PR:INSTITUTE Translational Genomics Research Institute PR:LAST_NAME Pirrotte PR:FIRST_NAME Patrick PR:ADDRESS 445 N 5TH STREET PR:EMAIL ims@tgen.org PR:PHONE 6023438454 PR:FUNDING_SOURCE NIH K12 grant no. 5K12CA001727–29, The Hyundai Hope on Wheels Young PR:FUNDING_SOURCE Investigator Award, NCI grant no.P30CA033572 PR:PROJECT_COMMENTS Study 3 of 4 PR:CONTRIBUTORS Lior Goldberg, Eric R. Haas, Jiaqi Wu, Bryan Garcia, Ryan Urak, Vibhuti Vyas, PR:CONTRIBUTORS Ruby Espinosa, Tamara Munoz, Shirley Bierkatz, Jamie R. Wagner, Jinny Paul, Mary PR:CONTRIBUTORS C. Clark, Dat Ngo, Ibrahim Aldoss, Stephen J. Forman & Xiuli Wang #STUDY ST:STUDY_TITLE Immunometabolic determinants of long-term response in leukapheresis T cells of ST:STUDY_TITLE the leukemia patients receiving CD19 CAR T cell therapy ST:STUDY_TYPE Untargeted metabolomics ST:STUDY_SUMMARY To understand the immunometabolic features of T cells conferring long-term ST:STUDY_SUMMARY response in leukemia patients, we investigated metabolome of leukapheresis T ST:STUDY_SUMMARY cells isolated from both long-term and short-term responders. We observed ST:STUDY_SUMMARY significant difference in the T cell metabolome of long-term vs short-term ST:STUDY_SUMMARY responders suggesting that metabolism in native Tcell has potential influence on ST:STUDY_SUMMARY antileukemic activity of CAR-T cells. ST:INSTITUTE Translational Genomics Research Institute ST:LAST_NAME Pirrotte ST:FIRST_NAME Patrick ST:ADDRESS 445 N 5TH STREET ST:EMAIL ims@tgen.org ST:PHONE 16023438454 ST:NUM_FEMALES 8 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male and female #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS LT10xTnmem495 LT10xTnmem495_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT10xTnmem495xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT10xTnmem495 LT10xTnmem495_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT10xTnmem495xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT11xTnmem188 LT11xTnmem188_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT11xTnmem188xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT11xTnmem188 LT11xTnmem188_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT11xTnmem188xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT12xTnmem518 LT12xTnmem518_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT12xTnmem518xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT12xTnmem518 LT12xTnmem518_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT12xTnmem518xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT13xTnmem544 LT13xTnmem544_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT13xTnmem544xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT13xTnmem544 LT13xTnmem544_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT13xTnmem544xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT14xTnmem552 LT14xTnmem552_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT14xTnmem552xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT14xTnmem552 LT14xTnmem552_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT14xTnmem552xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT1xTnmem209 LT1xTnmem209_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT1xTnmem209xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT1xTnmem209 LT1xTnmem209_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT1xTnmem209xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT2xTnmem466 LT2xTnmem466_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT2xTnmem466xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT2xTnmem466 LT2xTnmem466_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT2xTnmem466xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT3xTnmem431 LT3xTnmem431_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT3xTnmem431xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT3xTnmem431 LT3xTnmem431_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT3xTnmem431xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT4xTnmem379 LT4xTnmem379_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT4xTnmem379xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT4xTnmem379 LT4xTnmem379_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT4xTnmem379xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT5xTnmem293 LT5xTnmem293_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT5xTnmem293xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT5xTnmem293 LT5xTnmem293_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT5xTnmem293xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT6xTnmem543 LT6xTnmem543_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT6xTnmem543xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT6xTnmem543 LT6xTnmem543_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT6xTnmem543xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS Lt7xTnmem549 Lt7xTnmem549_HILICneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT7xTnmem549xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS Lt7xTnmem549 Lt7xTnmem549_HILICpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT7xTnmem549xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT8xTnmem245 LT8xTnmem245_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT8xTnmem245xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT8xTnmem245 LT8xTnmem245_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT8xTnmem245xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT9xTnmem505 LT9xTnmem505_HILICneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT9xTnmem505xHILICneg_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT9xTnmem505 LT9xTnmem505_HILICpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01B_R_LT9xTnmem505xHILICpos_28JUN24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT10xTnmem495 LT10xTnmem495_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT10xTnmem495xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT10xTnmem495 LT10xTnmem495_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT10xTnmem495xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT11xTnmem188 LT11xTnmem188_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT11xTnmem188xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT11xTnmem188 LT11xTnmem188_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT11xTnmem188xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT12xTnmem518 LT12xTnmem518_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT12xTnmem518xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT12xTnmem518 LT12xTnmem518_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT12xTnmem518xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT13xTnmem544 LT13xTnmem544_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT13xTnmem544xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT13xTnmem544 LT13xTnmem544_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT13xTnmem544xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT14xTnmem552 LT14xTnmem552_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT14xTnmem552xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT14xTnmem552 LT14xTnmem552_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT14xTnmem552xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT1xTnmem209 LT1xTnmem209_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT1xTnmem209xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT1xTnmem209 LT1xTnmem209_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT1xTnmem209xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT2xTnmem466 LT2xTnmem466_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT2xTnmem466xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT2xTnmem466 LT2xTnmem466_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT2xTnmem466xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT3xTnmem431 LT3xTnmem431_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT3xTnmem431xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT3xTnmem431 LT3xTnmem431_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT3xTnmem431xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT4xTnmem379 LT4xTnmem379_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT4xTnmem379xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT4xTnmem379 LT4xTnmem379_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT4xTnmem379xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT5xTnmem293 LT5xTnmem293_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT5xTnmem293xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT5xTnmem293 LT5xTnmem293_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT5xTnmem293xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT6xTnmem543 LT6xTnmem543_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT6xTnmem543xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT6xTnmem543 LT6xTnmem543_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT6xTnmem543xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS Lt7xTnmem549 Lt7xTnmem549_RPneg Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT7xTnmem549xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS Lt7xTnmem549 Lt7xTnmem549_RPpos Sample source:leukapheresis T cells | Outcome:Short-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT7xTnmem549xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT8xTnmem245 LT8xTnmem245_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT8xTnmem245xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT8xTnmem245 LT8xTnmem245_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT8xTnmem245xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT9xTnmem505 LT9xTnmem505_RPneg Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT9xTnmem505xRPneg_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw SUBJECT_SAMPLE_FACTORS LT9xTnmem505 LT9xTnmem505_RPpos Sample source:leukapheresis T cells | Outcome:Long-term responders RAW_FILE_NAME(File name)=160_0004_01C_R_LT9xTnmem505xRPpos_02JUL24_CELL_HUM_LUM_0000_07AB_20MIN_KP_KP_01.raw #COLLECTION CO:COLLECTION_SUMMARY De-identified clinical samples were collected from a subset of adult patients CO:COLLECTION_SUMMARY with relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL) who CO:COLLECTION_SUMMARY received investigational CD19-specific CAR T cells in a phase 1/2 clinical trial CO:COLLECTION_SUMMARY (NCT02146924) at City of Hope. Patient or donor's Tcells were collected by CO:COLLECTION_SUMMARY leukapheresis. The leukapheresis Tcells were used for metabolomics profiling. CO:SAMPLE_TYPE T-cells CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Tn/mem cells were enriched from leukapheresis from long-term and short-term TR:TREATMENT_SUMMARY responders were used for the study #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For leukapheresis T cells, metabolites were extracted using 0.4 mL of 0.4 mL of SP:SAMPLEPREP_SUMMARY solvent mixture (water: methanol: acetonitrile, 1:2.25:0.75, v/v/v) and 3 SP:SAMPLEPREP_SUMMARY freeze-thaw (30 seconds each) cycles. The resultant metabolite extracts from T SP:SAMPLEPREP_SUMMARY cells were recovered by centrifuging at 13,226 xg at 4°C for 15 min. For HILIC SP:SAMPLEPREP_SUMMARY chromatography, the extract was directly used LC-MS analysis. For reversed phase SP:SAMPLEPREP_SUMMARY chromatography, the extract was dried under vacuum centrifugation and the dried SP:SAMPLEPREP_SUMMARY extract was reconstituted in 20% Acetonitrile, 80% water and used used for LC-MS SP:SAMPLEPREP_SUMMARY analysis. SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Reversed Phase chromatography CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Thermo Hypersil Gold C18 (150 x 2.1mm, 1.9um) CH:SOLVENT_A 100% Water; 0.1% formic acid CH:SOLVENT_B 100% Acetonitrile; 0.1% formic acid CH:FLOW_GRADIENT 0% B for 1min, 0-50% B for 4 min, 50-98% B for 3 min, 98% B for 2 min, 0% B for CH:FLOW_GRADIENT 0.5min, 0% B for 5 min CH:FLOW_RATE 0.3 ml/min CH:COLUMN_TEMPERATURE 45 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap Fusion Lumos Tribrid MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Xcalibur Compound Discoverer 3.2 MS:MS_RESULTS_FILE ST003965_AN006521_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END